Lectin panning method: the prospective isolation of mouse neural progenitor cells by the attachment of cell surface N-glycans to Phaseolus vulgaris erythroagglutinating lectin-coated dishes.
Retrospective isolation of neural progenitor cells (NPCs) may cause deterioration of the phenotype during the long-term cultivation. Therefore, prospective isolation is essential for understanding the exact characteristics of intact NPCs in the brain. However, few suitable specific cell surface antigens on NPCs that could be used for their prospective isolation are available. The present study demonstrated that within 60 min after initial plating, embryonic day 12 (E12) brain cells firmly attach to several types of lectin-coated culture wells, including Phaseolus vulgaris erythroagglutinating lectin (E-PHA), concanavalin A (Con A) and wheat germ agglutinin (WGA). Approximately 80% of the cells isolated from E-PHA-coated wells expressed the nestin antigen, which is a specific intracellular marker for NPCs and the ratio of 5-bromo-2'-deoxyuridine (BrdU)-positive/nestin-positive cells to the cells attached on the E-PHA-coated wells was significantly higher than that of the cells attached on the wells coated with other adhesive substrates. The cells that were isolated from the E-PHA-coated wells continued to attach to the well for 1 week, while those isolated from Con A- and WGA-coated wells lost their attachment after 6 days and 1 day, respectively. Furthermore, the cells isolated from the E-PHA-coated wells grew quite satisfactorily and formed numerous attached neurospheres. Their growth rate was almost equal to that observed in suspension cultures. These results indicate that the lectin panning method enables the prospective, quick and easy isolation of mouse NPCs without requiring a fluorescence-activated cell sorter (FACS) system.
Hamanoue
M
M
Department of Physiology, Toho University School of Medicine, 5-21-16 Ohmori-nishi, Ohta-ku, Tokyo 143-8540, Japan. hamanoue-ns@umin.ac.jp
Sato
K
K
Takamatsu
K
K
D000704:Analysis of Variance; D000818:Animals; D001921:Brain / Q000166:cytology; D001973:Bromodeoxyuridine / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects* / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000502:physiology; D005260:Female; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010835:Phytohemagglutinins / Q000494:pharmacology*; D011247:Pregnancy; D014909:Wheat Germ Agglutinins / Q000494:pharmacology
D009419:Nerve Tissue Proteins; D010835:Phytohemagglutinins; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; D001973:Bromodeoxyuridine